Screening for resistance to Elsinoe ampelina (de Bary) Shear, causal agent of grape anthracnose, in grapevine seedlings is commonly conducted by natural infection over 3 to 4 years in the vineyard. The objective of this research was to develop a greenhouse screening method for selecting grapevine seedlings with resistance to anthracnose. Spores of E. ampelina were obtained from 3- to 4-week-old cultures on potato dextrose agar. Inoculum concentrations ranging from 1.3 × 103 to 1.3 × 107E. ampelina conidia per mL were evaluated and 106 conidia/mL was optimum. The time of incubation of seedlings in a moist chamber after inoculation varied from 24 to 120 hours with 24 to 72 hours resulting in good symptom development. Temperatures in the moist chamber from 16 to 32 °C were evaluated and the most consistent results were obtained at 20 to 28 °C. The most effective method for selecting anthracnose-resistant grape seedlings in the two-to-three true-leaf stage was misting the seedlings with a suspension containing 106 conidia/mL in water and placing the inoculated seedlings in a moist chamber at 24 °C for 48 hours, followed by 8 days on a greenhouse bench. Resistant seedlings from the greenhouse screening (those with <10 foliar lesions) were transplanted into the vineyard and found to be resistant to anthracnose infection under rainy, humid conditions. This greenhouse procedure for selecting grapevine cultivars and breeding lines with resistance to anthracnose is accurate, economical, and labor-saving.